Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Jul 1;109(1):331–339. doi: 10.1083/jcb.109.1.331

Mechano-chemical control of human endothelium orientation and size

PMCID: PMC2115455  PMID: 2545727

Abstract

Human umbilical vein endothelial cells (EC) were grown on elastic silicone membranes subjected to cyclic stretch, simulating arterial wall motion. Stretching conditions (20% amplitude, 52 cycle/min) stimulated stress fiber formation and their orientation transversely to the strain direction. Cell bodies aligned along the same axis after the actin cytoskeleton. EC orientation response was inhibited by the adenylate cyclase activator, forskolin (10(-5) M), which caused stress fiber disassembly and the redistribution of F-actin to the cortical cytoplasm. Preoriented EC depleted of stress fibers by forskolin treatment retained their aligned state. Thus, stress fibers are essential for the process of EC orientation induced by repeated strain, but not for the maintenance of EC orientation. The monolayer formed by EC grown to confluence in conditions of intermittent strain consisted of uniform elongated cells and was resistant to deformation. In contrast, the monolayer assembled in stationary conditions was less compliant and exposed local denudations on initiation of stretching. When stretched in the presence of 10(-5) M forskolin it rapidly (3-4 h) reestablished integrity but gained a heterogeneous appearance since denuded areas were covered by giant cells. The protective effect of forskolin was because of the stimulation of EC spreading. This feature of forskolin was demonstrated while studying its action on EC spreading and repair of a scratched EC monolayer in conventional culture. Thus mechanical deformation and adenylate cyclase activity may be important factors in the control of endothelium morphology in human arteries.

Full Text

The Full Text of this article is available as a PDF (4.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adelstein R. S., Conti M. A., Hathaway D. R., Klee C. B. Phosphorylation of smooth muscle myosin light chain kinase by the catalytic subunit of adenosine 3': 5'-monophosphate-dependent protein kinase. J Biol Chem. 1978 Dec 10;253(23):8347–8350. [PubMed] [Google Scholar]
  2. Antonov A. S., Lukashev M. E., Romanov Y. A., Tkachuk V. A., Repin V. S., Smirnov V. N. Morphological alterations in endothelial cells from human aorta and umbilical vein induced by forskolin and phorbol 12-myristate 13-acetate: a synergistic action of adenylate cyclase and protein kinase C activators. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9704–9708. doi: 10.1073/pnas.83.24.9704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Antonov A. S., Nikolaeva M. A., Klueva T. S., Romanov YuA, Babaev V. R., Bystrevskaya V. B., Perov N. A., Repin V. S., Smirnov V. N. Primary culture of endothelial cells from atherosclerotic human aorta. Part 1. Identification, morphological and ultrastructural characteristics of two endothelial cell subpopulations. Atherosclerosis. 1986 Jan;59(1):1–19. doi: 10.1016/0021-9150(86)90027-4. [DOI] [PubMed] [Google Scholar]
  4. Booyse F. M., Osikowicz G., Quarfoot A. J. Effects of chronic oral consumption of nicotine on the rabbit aortic endothelium. Am J Pathol. 1981 Feb;102(2):229–238. [PMC free article] [PubMed] [Google Scholar]
  5. Booyse F. M., Sedlak B. J., Rafelson M. E., Jr Culture of arterial endothelial cells: characterization and growth of bovine aortic cells. Thromb Diath Haemorrh. 1975 Dec 15;34(3):825–839. [PubMed] [Google Scholar]
  6. Buck R. C. Behavior of vascular smooth muscle cells during repeated stretching of the substratum in vitro. Atherosclerosis. 1983 Feb;46(2):217–223. doi: 10.1016/0021-9150(83)90112-0. [DOI] [PubMed] [Google Scholar]
  7. Buck R. C. Reorientation response of cells to repeated stretch and recoil of the substratum. Exp Cell Res. 1980 Jun;127(2):470–474. doi: 10.1016/0014-4827(80)90456-5. [DOI] [PubMed] [Google Scholar]
  8. Dartsch P. C., Hämmerle H., Betz E. Orientation of cultured arterial smooth muscle cells growing on cyclically stretched substrates. Acta Anat (Basel) 1986;125(2):108–113. doi: 10.1159/000146146. [DOI] [PubMed] [Google Scholar]
  9. Dartsch P. C., Hämmerle H. Orientation response of arterial smooth muscle cells to mechanical stimulation. Eur J Cell Biol. 1986 Aug;41(2):339–346. [PubMed] [Google Scholar]
  10. Dewey C. F., Jr, Bussolari S. R., Gimbrone M. A., Jr, Davies P. F. The dynamic response of vascular endothelial cells to fluid shear stress. J Biomech Eng. 1981 Aug;103(3):177–185. doi: 10.1115/1.3138276. [DOI] [PubMed] [Google Scholar]
  11. Franke R. P., Gräfe M., Schnittler H., Seiffge D., Mittermayer C., Drenckhahn D. Induction of human vascular endothelial stress fibres by fluid shear stress. Nature. 1984 Feb 16;307(5952):648–649. doi: 10.1038/307648a0. [DOI] [PubMed] [Google Scholar]
  12. Heggeness M. H., Wang K., Singer S. J. Intracellular distributions of mechanochemical proteins in cultured fibroblasts. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3883–3887. doi: 10.1073/pnas.74.9.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herman I. M., Brant A. M., Warty V. S., Bonaccorso J., Klein E. C., Kormos R. L., Borovetz H. S. Hemodynamics and the vascular endothelial cytoskeleton. J Cell Biol. 1987 Jul;105(1):291–302. doi: 10.1083/jcb.105.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holzapfel G., Wehland J., Weber K. Calcium control of actin-myosin based contraction in triton models of mouse 3T3 fibroblasts is mediated by the myosin light chain kinase (MLCK)-calmodulin complex. Exp Cell Res. 1983 Oct;148(1):117–126. doi: 10.1016/0014-4827(83)90192-1. [DOI] [PubMed] [Google Scholar]
  15. Ives C. L., Eskin S. G., McIntire L. V. Mechanical effects on endothelial cell morphology: in vitro assessment. In Vitro Cell Dev Biol. 1986 Sep;22(9):500–507. doi: 10.1007/BF02621134. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lewis J. C., Taylor R. G., Jones N. D., St Clair R. W., Cornhill J. F. Endothelial surface characteristics in pigeon coronary artery atherosclerosis. I. Cellular alterations during the initial stages of dietary cholesterol challenge. Lab Invest. 1982 Feb;46(2):123–138. [PubMed] [Google Scholar]
  18. Repin V. S., Dolgov V. V., Zaikina O. E., Novikov I. D., Antonov A. S., Nikolaeva M. A., Smirnov V. N. Heterogeneity of endothelium in human aorta. A quantitative analysis by scanning electron microscopy. Atherosclerosis. 1984 Jan;50(1):35–52. doi: 10.1016/0021-9150(84)90006-6. [DOI] [PubMed] [Google Scholar]
  19. Scholey J. M., Taylor K. A., Kendrick-Jones J. Regulation of non-muscle myosin assembly by calmodulin-dependent light chain kinase. Nature. 1980 Sep 18;287(5779):233–235. doi: 10.1038/287233a0. [DOI] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Verkhovsky A. B., Surgucheva I. G., Svitkina T. M., Tint I. S., Gelfand V. I. Organization of stress fibers in cultured fibroblasts after extraction of actin with bovine brain gelsolin-like protein. Exp Cell Res. 1987 Nov;173(1):244–255. doi: 10.1016/0014-4827(87)90349-1. [DOI] [PubMed] [Google Scholar]
  22. Wong A. J., Pollard T. D., Herman I. M. Actin filament stress fibers in vascular endothelial cells in vivo. Science. 1983 Feb 18;219(4586):867–869. doi: 10.1126/science.6681677. [DOI] [PubMed] [Google Scholar]
  23. Wong M. K., Gotlieb A. I. Endothelial cell monolayer integrity. I. Characterization of dense peripheral band of microfilaments. Arteriosclerosis. 1986 Mar-Apr;6(2):212–219. doi: 10.1161/01.atv.6.2.212. [DOI] [PubMed] [Google Scholar]
  24. Zand T., Underwood J. M., Nunnari J. J., Majno G., Joris I. Endothelium and "silver lines". An electron microscopic study. Virchows Arch A Pathol Anat Histol. 1982;395(2):133–144. doi: 10.1007/BF00429607. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES